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Density dependent regulation of inflammatory responses in macrophages

Alun Vaughan-Jackson1, Szymon Stodolak1, Kourosh H Ebrahimi2

  • 1James & Lillian Martin Centre, Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom.

Frontiers in Immunology
|January 2, 2023
PubMed

Insights

Macrophage crowding in lab cultures significantly alters their inflammatory responses. This density-dependent effect, observed across various macrophage types, is crucial for interpreting in vitro research.

Area of Science:

  • Immunology
  • Cell Biology
  • Stem Cell Research

Background:

  • Macrophage distribution is tightly regulated in vivo.
  • The impact of macrophage density during in vitro culture is understudied.
  • Understanding in vitro conditions is vital for accurate biological interpretation.

Purpose of the Study:

  • To investigate how increasing macrophage culture density affects their morphology and phenotype.
  • To determine the influence of density on inflammatory cytokine and chemokine secretion.
  • To assess if density-dependent effects are conserved across different macrophage models.

Main Methods:

  • Utilized human induced pluripotent stem cell (iPSC)-derived macrophages.
  • Compared macrophage behavior at varying culture densities.
  • Analyzed morphology, phenotype, and inflammatory mediator secretion.
  • Tested density effects in blood monocyte-derived macrophages, THP-1 cells, and iPSC-derived microglia.

Main Results:

  • Increasing macrophage density alters cell morphology and phenotype.
  • Culture density significantly impacts inflammatory cytokine and chemokine secretion in both resting and activated states.
  • Density-dependent inflammatory modulation is observed in iPSC-derived macrophages, blood monocyte-derived macrophages, THP-1 cells, and iPSC-derived microglia.
  • A transferable soluble factor appears to mediate density-dependent effects.

Conclusions:

  • Cell plating density is a critical, often overlooked, factor in in vitro macrophage research.
  • Density-dependent regulation of the macrophage inflammatory state has broad implications.
  • Further research is needed to elucidate the precise mechanism of density-dependent regulation.
Keywords:
densitydifferentiationinflammationmacrophagemicrogliastem cell

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